Stereospecific carbonyl reductases

Abstract

Stereospecific carbonyl reductases SCR1, SCR2, and SCR3 are described herein as are nucleotide sequences that encode these reductases. These stereospecific carbonyl reductases have anti-Prelog selectivity and have specificities that are useful for fine biochemical synthesis.

We claim: 
     
       1. A method of reducing a carbonyl substrate, comprising contacting the substrate with a purified polypeptide having carbonyl reductase activity, the sequence of which comprises an amino acid sequence that has at least 80% sequence identity to SEQ ID NO:4, at least 95% sequence identity to SEQ ID NO:3 or at least 90% sequence identity to SEQ ID NO:2, in conditions suitable to catalyze the reduction of the carbonyl substrate. 
     
     
       2. The method of  claim 1 , wherein the reduction takes place in the presence of a coenzyme. 
     
     
       3. The method of  claim 2 , wherein the coenzyme is NADPH. 
     
     
       4. The method of  claim 1 , wherein the carbonyl substrate comprises an α-ketoester, a β-ketoester, an aryl ketone or an aliphatic ketone. 
     
     
       5. The method of  claim 4 , wherein the carbonyl substrate comprises an α-ketoester. 
     
     
       6. The method of  claim 5 , wherein the α-ketoester is methyl pyruvate, methyl phenylglyoxylate, ethyl pyruvate or ethyl benzoylformate. 
     
     
       7. The method of  claim 4 , wherein the carbonyl substrate comprises a β-ketoester. 
     
     
       8. The method of claim of  claim 7 , wherein the β-ketoester is ethyl trifluoroacetoacetate, methyl acetoacetate, methyl 3-oxovalerate, methyl 4-fluorobenzoylacetate, ethyl acetoacetate, ethyl 3-oxovalerate, ethyl 4-chloroacetoacetate, ethyl benzoylacetate, or ethyl 3,4-dimethoxybenzoylacetate. 
     
     
       9. The method of  claim 4 , wherein the carbonyl substrate comprises an aryl ketone. 
     
     
       10. The method of  claim 9 , wherein the aryl ketone is 2-hydroxyacetophenone, or a derivative thereof. 
     
     
       11. The method of  claim 10 , wherein the aryl ketone is 2′-chloro-2-hydroxyacetophenone, 3′-chloro-2-hydroxyacetophenone, 4′-chloro-2-hydroxyacetophenone or 4′-methoxy-2-hydroxyacetophenone. 
     
     
       12. The method of  claim 4 , wherein the carbonyl substrate comprises an aliphatic ketone. 
     
     
       13. The method of  claim 12 , wherein the aliphatic ketone is 2-butanone, 2-pentanone, 2-hexanone, 2-heptanone, 2-octanone or 2-methyl-3-pentanone. 
     
     
       14. The method of  claim 1 , wherein the carbonyl substrate is ethyl 4-chloro-3-oxobutyrate. 
     
     
       15. The method of  claim 1 , wherein the reduction takes place at pH ranging from 5.0 to 6.0. 
     
     
       16. The method of  claim 1 , wherein the amino acid sequence has at least 85% sequence identity to SEQ ID NO:4. 
     
     
       17. The method of  claim 1 , wherein the amino acid sequence has at least 90% sequence identity to SEQ ID NO:4. 
     
     
       18. The method of  claim 1 , wherein the amino acid sequence has at least 95% sequence identity to SEQ ID NO:4. 
     
     
       19. The method of  claim 1 , wherein the amino acid sequence has at least 99% sequence identity to SEQ ID NO:3. 
     
     
       20. The method of  claim 1 , wherein the amino acid sequence has at least 99% sequence identity to SEQ ID NO:2. 
     
     
       21. The method of  claim 1 , wherein the amino acid sequence has at least 95% sequence identity to SEQ ID NO:2. 
     
     
       22. The method of  claim 1 , wherein the amino acid sequence comprises SEQ ID NO:4, SEQ ID NO:3 or SEQ ID NO:2. 
     
     
       23. The method of  claim 1 , wherein the amino acid sequence consists of SEQ ID NO:4, SEQ ID NO:3 or SEQ ID NO:2. 
     
     
       24. The method of  claim 1 , wherein the amino acid sequence comprises SEQ ID NO:9, SEQ ID NO:10 and/or SEQ ID NO:11.